Evaluation of one-step RT-PCR multiplex assay for body fluid identification.

Qinrui Yang, Baonian Liu, Yuxiang Zhou, Yining Yao, Zhihan Zhou, Hui Li, Chengchen Shao, Kuan Sun, Hongmei Xu, Qiqun Tang, Yiwen Shen, Jianhui Xie

International journal of legal medicine 2021 Sep

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The discrimination of body fluid stains provides crucial evidence during the investigation of criminal cases. Previous studies have demonstrated the practical value of mRNA profiling in body fluid identification. Conventional strategy of mRNA profiling entails reverse transcription and PCR amplification in two separate procedures with different buffer systems. In this study, we subjected the one-step multiplex reverse transcription PCR strategy to mRNA profiling with the inclusion of the same 18 tissue-specific biomarkers in the F18plex system targeting peripheral blood, menstrual blood, vaginal secretion, saliva, semen, and urine. The Qiagen OneStep RT-PCR kit and Titanium One-Step RT-PCR kit were applied to multiplex construction, while reproducible profiling results were obtained with both kits. Compared to the F18plex system, similar expression profiles of biomarkers were obtained in targeted tissues, while expected cross-reaction was observed in non-targeted body fluids. However, CYP2B7P1 and SPINK5 were detected in menstrual blood samples, which was not observed using the F18plex system. Full-profiling results were obtained in all samples using 0.1 ng peripheral blood and semen RNA, and 1 ng menstrual blood, vaginal secretion, saliva, and urine RNA. In conclusion, the application of one-step mRNA profiling strategy could be a reliable and economical method for the simplified, specific, and simultaneous analysis of tissue-specific biomarkers for the discrimination of body fluid origin. © 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.

Citation

Qinrui Yang, Baonian Liu, Yuxiang Zhou, Yining Yao, Zhihan Zhou, Hui Li, Chengchen Shao, Kuan Sun, Hongmei Xu, Qiqun Tang, Yiwen Shen, Jianhui Xie. Evaluation of one-step RT-PCR multiplex assay for body fluid identification. International journal of legal medicine. 2021 Sep;135(5):1727-1735